A simple and reliable PCR-restriction fragment length polymorphism assay to identify Candida albicans and its closely related Candida dubliniensis
Braz. j. microbiol
; 43(3): 873-879, July-Sept. 2012. ilus, tab
Article
en En
| LILACS
| ID: lil-656647
Biblioteca responsable:
BR32.1
ABSTRACT
Candida dubliniensis is an emerging pathogen capable of causing superficial as well as systemic infections. Due to its close similarity to C. albcians, conventional methods based on phenotypic traits are not always reliable in identification of C. dubliniensis. In this study, we developed a PCR-restriction fragment length polymorphism (RFLP) assay to identify and discriminate between the two closely related species. The D1/D2 region of 28S rDNA was amplified by PCR and enzymatically digested by ApaI and BsiEI respectively. PCR products of both species were digested into two fragments by ApaI, but those of other yeast species were undigested. BsiEI cut the PCR products of C. albicans into two fragments but not those of C. dubliniensis. Thus two species were differentiated. We evaluated 10 reference strains representing 10 yeast species, among which C. albicans and C. dubliniensis were successfully identified. A total of 56 phenotypically characterized clinical isolates (42 C. albicans isolates and 14 C. dubliniensis isolates) were also investigated for intra-species variability. All tested isolates produced identical RFLP patterns to their respective reference strains except one initially misidentified isolate. Our method offers a simple, rapid and reliable molecular method for the identification of C. albicans and C. dubliniensis.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
LILACS
Asunto principal:
Fenotipo
/
Polimorfismo Genético
/
Candida albicans
/
Candidiasis
/
Reacción en Cadena de la Polimerasa
Tipo de estudio:
Evaluation_studies
Límite:
Humans
Idioma:
En
Revista:
Braz. j. microbiol
Asunto de la revista:
MICROBIOLOGIA
Año:
2012
Tipo del documento:
Article
País de afiliación:
China